title: Silencing of GLS and overexpression of GLS2 genes cooperate in decreasing the proliferation and viability of glioblastoma cells.
abstract: Glutamine (Gln) metabolism, initiated by its degradation by glutaminases (GA), is elevated in neoplastic cells and tissues. In malignant glia-derived tumors, GA isoforms, KGA and GAC, coded by the GLS gene, are overexpressed, whereas the GLS2-coded GAB and LGA isoforms, are hardly detectable in there. Our previous study revealed that transfection of T98G glioblastoma cells with GAB reduced cell proliferation and migration, by a yet unknown mechanism not related to Gln degradation. The question arose how simultaneous overexpression of GAB and inhibition of KGA would affect glioblastoma cell growth. Here, we used siRNA to silence the expression of Gls in T98G cells which were or were not stably transfected with GAB (TGAB cells). In both T98G and TGAB cell lines, silencing of Gls with siRNAs targeted at different sequences decreased cell viability and proliferation in a different, sequence-dependent degree, and the observed decreases were in either cell line highly correlated with increase of intracellular Gln (r > 0.9), a parameter manifesting decreased Gln degradation. The results show that combination of negative modulation of GA isoforms arising from GLS gene with the introduction of the GLS2 gene product, GAB, may in the future provide a useful means to curb glioblastoma growth in situ. At the same time, the results underscore the critical role of Gln degradation mediated by KGA in the manifestations of aggressive glial tumor phenotype.
authors.affiliation: Department of Neurotoxicology, Mossakowski Medical Research Centre Polish Academy of Sciences, 5 Pawińskiego Str., 02-106, Warsaw, Poland, mszeliga@imdik.pan.pl.
authors.forename: Monika
authors.initials: M
authors.lastname: Szeliga
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Bogacińska-Karaś
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Różycka
authors.forename: Wojciech
authors.initials: W
authors.lastname: Hilgier
authors.forename: Javier
authors.initials: J
authors.lastname: Marquez
authors.forename: Jan
authors.initials: J
authors.lastname: Albrecht
mesh_terms: D015153:Blotting, Western; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival / Q000235:genetics; D020868:Gene Silencing; D005909:Glioblastoma / Q000201:enzymology* / Q000235:genetics*; D005972:Glutaminase / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
chemical_list: D007527:Isoenzymes; D034741:RNA, Small Interfering; C586504:GLS2 protein, human; D005972:Glutaminase